Zang-Si-Wei and its core constituent ellagic acid alleviate LPS-induced acute lung injury by modulating PI3K/AKT signaling and NLRP3-associated pyroptosis

Background Acute lung injury (ALI) is a life-threatening respiratory condition characterized by uncontrolled pulmonary inflammation and epithelial barrier dysfunction. Zang-Si-Wei Qingfei Mixture (ZSW) is a promising Tibetan herbal formula used clinically to alleviate pulmonary symptoms, yet its pharmacological targets and therapeutic pathways in ALI have not been fully explored. This study aims to identify the bioactive constituents of ZSW and elucidate their protective and therapeutic mechanisms in ALI. Methods ZSW’s components were identified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) previously and network pharmacology. Target affinity was evaluated using molecular docking and molecular dynamics simulations. In vivo , C57BL/6J mice with lipopolysaccharide (LPS)-induced ALI were used to evaluate the pretreatment and post-treatment effects of ZSW and its core component, Ellagic Acid (EA). In vitro , MLE-12 cells and the PI3K/mTOR dual inhibitor BEZ235 were utilized to investigate the role of PI3K/AKT signaling and NLRP3 inflammasome activation and pyroptosis. Results EA was identified as ZSW’s core bioactive constituent, demonstrating high binding stability with AKT1, PIK3CA, and PIK3R1. In vivo , ZSW and EA significantly attenuated lung histological damage, edema, and alveolar-capillary hyperpermeability. Both markedly reduced pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) in bronchoalveolar lavage fluid. Mechanistically, ZSW and EA inhibited PI3K and AKT phosphorylation and were accompanied by decreased NLRP3 inflammasome-associated proteins (decreased NLRP3, ASC, and cleaved Caspase-1) and GSDMD-mediated pyroptosis (decreased GSDMD-N and lactate dehydrogenase release). Notably, the 6-h post-LPS therapeutic model showed that both treatments effectively reversed established lung injury. Conclusion ZSW and EA exert anti-inflammatory and lung-protective effects, accompanied by modulation of PI3K/AKT signaling and suppression of NLRP3 inflammasome-associated pyroptosis. This provides robust scientific evidence for ZSW’s clinical application and highlights EA as a promising therapeutic candidate for acute inflammatory lung diseases.